A novel artificial peroxidase (AP) with highly catalytic efficiency was designed using hexadecyl trimethyl ammonium bromide (CTAB, 3 mM) nano-micelles and bovine heart cytochrome c (Cyt c, 0.5 µM) in 100 mM, pH 8.0 phosphate buffer at 25°C. The catalytic rate (kcat) and Michaelis-Menten (Km) of the AP were determined to be 0.311 ± 0.013 s−1 and 8.64 ± 0.6 µM. The catalytic efficiency was 0.0360 ± 0.0020 µM−1s−1 (about 50% the efficiency of native horseradish peroxidase). The Ultraviolet–visible spectrophotometer and Circular Dichroism techniques were applied to study the properties of the CTAB-Cyt c nano-micelle. Designed AP can be applied instead of native horseradish peroxidase.